LOCUS       sequence-206124-       12886 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dCas9 and DNMT WT on C terminus, 4 NLS, driven by pGK promoter, with
            P2A site and PURO gene, within a lentiviral transfer backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12886)
  AUTHORS   Pflueger C, Tan D, Swain T, Nguyen TV, Pflueger J, Nefzger C, Polo 
            JM, Ford E, Lister R
  TITLE     A modular dCas9-SunTag DNMT3A epigenome editing system overcomes 
            pervasive off-target activity of direct fusion dCas9-DNMT3A 
            constructs.
  JOURNAL   Genome Res. 2018 Jun 15. pii: gr.233049.117. doi: 
            10.1101/gr.233049.117.
  PUBMED    29907613
REFERENCE   2  (bases 1 to 12886)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genome Res.
            2018 Jun 15. pii: gr.233049.117. doi: 10.1101/gr.233049.117."
COMMENT     Sequence Label: LLP185 pLVP-dCas9-DNMT3a V2
FEATURES             Location/Qualifiers
     source          1..12886
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2028..2143
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        2214..2713
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2249..2272)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2646..2665
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2737..2757
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2758..2784
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2788..2814
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2818..2844
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(2878..2904,2905..2907,2908..5391,5392..5397,5398..5463,
                     5464..5466,5467..6981)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(2878..2904,2905..2907,2908..5394,5395..5397,
                     5398..6981)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             2878..6981
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7018..7038
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7078..7098
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7120..8052
                     /codon_start=1
                     /product="catalytic domain of DNA methyltransferase 3A"
                     /label=DNMT3A catalytic domain
                     /translation="PSRLQMFFANNHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATG
                     LLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVI
                     GGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVS
                     DKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGR
                     IAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSR
                     LARQRLLGRSWSVPVIRHLFAPLKEYFACV"
     CDS             8152..8172
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8182..8238
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             8239..8838
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(8239..8258)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8735..8755
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8855..9443
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8908..8928)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9326..9337)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9650..10283
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10411..10427)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10411..10427)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10424..10446)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10435..10451
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(10459..10465,10466..10483,10484..10489))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10504..10525
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10642..10659)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10813..11401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10893..10912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11572..12363,12364..12432))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     12195..12214
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12433..12537)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    12585..12719
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12622..12641)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12676..12695
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgatgaggcc ctttcgtctt cactcgagtt accgggtagg
     2221 ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg
     2281 gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc
     2341 ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt
     2401 cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac
     2461 tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag
     2521 cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc
     2581 cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg
     2641 cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc
     2701 tccgggcctt tcgacctgca gcccaagctt accatgccaa agaagaagcg gaaggtctac
     2761 ccctatgacg tgcctgatta cgccgcttat ccttacgatg tgcccgacta tgcagcctac
     2821 ccatacgatg tgcccgacta cgccgcaatg gccggtatcc acggagtccc agcagccatg
     2881 gacaagaagt actccattgg gctcgctatc ggcacaaaca gcgtcggctg ggccgtcatt
     2941 acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc tgggcaatac cgatcgccac
     3001 agcataaaga agaacctcat tggcgccctc ctgttcgact ccggggagac ggccgaagcc
     3061 acgcggctca aaagaacagc acggcgcaga tatacccgca gaaagaatcg gatctgctac
     3121 ctgcaggaga tctttagtaa tgagatggct aaggtggatg actctttctt ccataggctg
     3181 gaggagtcct ttttggtgga ggaggataaa aagcacgagc gccacccaat ctttggcaat
     3241 atcgtggacg aggtggcgta ccatgaaaag tacccaacca tatatcatct gaggaagaag
     3301 cttgtagaca gtactgataa ggctgacttg cggttgatct atctcgcgct ggcgcatatg
     3361 atcaaatttc ggggacactt cctcatcgag ggggacctga acccagacaa cagcgatgtc
     3421 gacaaactct ttatccaact ggttcagact tacaatcagc ttttcgaaga gaacccgatc
     3481 aacgcatccg gagttgacgc caaagcaatc ctgagcgcta ggctgtccaa atcccggcgg
     3541 ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga acggcctgtt tggtaatctt
     3601 atcgccctgt cactcgggct gacccccaac tttaaatcta acttcgacct ggccgaagat
     3661 gccaagcttc aactgagcaa agacacctac gatgatgatc tcgacaatct gctggcccag
     3721 atcggcgacc agtacgcaga cctttttttg gcggcaaaga acctgtcaga cgccattctg
     3781 ctgagtgata ttctgcgagt gaacacggag atcaccaaag ctccgctgag cgctagtatg
     3841 atcaagcgct atgatgagca ccaccaagac ttgactttgc tgaaggccct tgtcagacag
     3901 caactgcctg agaagtacaa ggaaattttc ttcgatcagt ctaaaaatgg ctacgccgga
     3961 tacattgacg gcggagcaag ccaggaggaa ttttacaaat ttattaagcc catcttggaa
     4021 aaaatggacg gcaccgagga gctgctggta aagcttaaca gagaagatct gttgcgcaaa
     4081 cagcgcactt tcgacaatgg aagcatcccc caccagattc acctgggcga actgcacgct
     4141 atcctcaggc ggcaagagga tttctacccc tttttgaaag ataacaggga aaagattgag
     4201 aaaatcctca catttcggat accctactat gtgggccccc tcgcccgggg aaattccaga
     4261 ttcgcgtgga tgactcgcaa atcagaagag accatcactc cctggaactt cgaggaagtc
     4321 gtggataagg gggcctctgc ccagtccttc atcgaaagga tgactaactt tgataaaaat
     4381 ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt acgagtactt cacagtttat
     4441 aacgagctca ccaaggtcaa atacgtcaca gaagggatga gaaagccagc attcctgtct
     4501 ggagagcaga agaaagctat cgtggacctc ctcttcaaga cgaaccggaa agttaccgtg
     4561 aaacagctca aagaagacta tttcaaaaag attgaatgtt tcgactctgt tgaaatcagc
     4621 ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc acgatctcct gaaaatcatt
     4681 aaagacaagg acttcctgga caatgaggag aacgaggaca ttcttgagga cattgtcctc
     4741 acccttacgt tgtttgaaga tagggagatg attgaagaac gcttgaaaac ttacgctcat
     4801 ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc gatatacagg atgggggcgg
     4861 ctgtcaagaa aactgatcaa tgggatccga gacaagcaga gtggaaagac aatcctggat
     4921 tttcttaagt ccgatggatt tgccaacagg aacttcatgc agttgatcca tgatgactct
     4981 ctcaccttta aggaggacat ccagaaagca caagtttctg gccaggggga cagtcttcac
     5041 gagcacatcg ctaatcttgc aggtagccca gctatcaaaa agggaatact gcagaccgtt
     5101 aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata agcccgagaa tatcgttatc
     5161 gagatggccc gagagaacca aactacccag aagggacaga agaacagtag ggaaaggatg
     5221 aagaggattg aagagggtat aaaagaactg gggtcccaaa tccttaagga acacccagtt
     5281 gaaaacaccc agcttcagaa tgagaagctc tacctgtact acctgcagaa cggcagggac
     5341 atgtacgtgg atcaggaact ggacatcaat cggctctccg actacgacgt ggatgcaatc
     5401 gtgccccagt cttttctcaa agatgattct attgataata aagtgttgac aagatccgat
     5461 aaaaatagag ggaagagtga taacgtcccc tcagaagaag ttgtcaagaa aatgaaaaat
     5521 tattggcggc agctgctgaa cgccaaactg atcacacaac ggaagttcga taatctgact
     5581 aaggctgaac gaggtggcct gtctgagttg gataaagccg gcttcatcaa aaggcagctt
     5641 gttgagacac gccagatcac caagcacgtg gcccaaattc tcgattcacg catgaacacc
     5701 aagtacgatg aaaatgacaa actgattcga gaggtgaaag ttattactct gaagtctaag
     5761 ctggtctcag atttcagaaa ggactttcag ttttataagg tgagagagat caacaattac
     5821 caccatgcgc atgatgccta cctgaatgca gtggtaggca ctgcacttat caaaaaatat
     5881 cccaagcttg aatctgaatt tgtttacgga gactataaag tgtacgatgt taggaaaatg
     5941 atcgcaaagt ctgagcagga aataggcaag gccaccgcta agtacttctt ttacagcaat
     6001 attatgaatt ttttcaagac cgagattaca ctggccaatg gagagattcg gaagcgacca
     6061 cttatcgaaa caaacggaga aacaggagaa atcgtgtggg acaagggtag ggatttcgcg
     6121 acagtccgga aggtcctgtc catgccgcag gtgaacatcg ttaaaaagac cgaagtacag
     6181 accggaggct tctccaagga aagtatcctc ccgaaaagga acagcgacaa gctgatcgca
     6241 cgcaaaaaag attgggaccc caagaaatac ggcggattcg attctcctac agtcgcttac
     6301 agtgtactgg ttgtggccaa agtggagaaa gggaagtcta aaaaactcaa aagcgtcaag
     6361 gaactgctgg gcatcacaat catggagcga tcaagcttcg aaaaaaaccc catcgacttt
     6421 ctcgaggcga aaggatataa agaggtcaaa aaagacctca tcattaagct tcccaagtac
     6481 tctctctttg agcttgaaaa cggccggaaa cgaatgctcg ctagtgcggg cgagctgcag
     6541 aaaggtaacg agctggcact gccctctaaa tacgttaatt tcttgtatct ggccagccac
     6601 tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga agcagctgtt cgtggaacaa
     6661 cacaaacact accttgatga gatcatcgag caaataagcg aattctccaa aagagtgatc
     6721 ctcgccgacg ctaacctcga taaggtgctt tctgcttaca ataagcacag ggataagccc
     6781 atcagggagc aggcagaaaa cattatccac ttgtttactc tgaccaactt gggcgcgcct
     6841 gcagccttca agtacttcga caccaccata gacagaaagc ggtacacctc tacaaaggag
     6901 gtcctggacg ccacactgat tcatcagtca attacggggc tctatgaaac aagaatcgac
     6961 ctctctcagc tcggtggaga cagcagggct gacgggggag gatcaggggg agggtccccc
     7021 aaaaagaaac gcaaagtgac cggcaagaaa accagcggcc aggccggaca ggcctctcct
     7081 aagaaaaaaa gaaaagtggg acgcgctggc ggcggaagcc cctcccggct ccagatgttc
     7141 ttcgctaata accacgacca ggaatttgac cctccaaagg tttacccacc tgtcccagct
     7201 gagaagagga agcccatccg ggtgctgtct ctctttgatg gaatcgctac agggctcctg
     7261 gtgctgaagg acttgggcat tcaggtggac cgctacattg cctcggaggt gtgtgaggac
     7321 tccatcacgg tgggcatggt gcggcaccag gggaagatca tgtacgtcgg ggacgtccgc
     7381 agcgtcacac agaagcatat ccaggagtgg ggcccattcg atctggtgat tgggggcagt
     7441 ccctgcaatg acctctccat cgtcaaccct gctcgcaagg gcctctacga gggcactggc
     7501 cggctcttct ttgagttcta ccgcctcctg catgatgcgc ggcccaagga gggagatgat
     7561 cgccccttct tctggctctt tgagaatgtg gtggccatgg gcgttagtga caagagggac
     7621 atctcgcgat ttctcgagtc caaccctgtg atgattgatg ccaaagaagt gtcagctgca
     7681 cacagggccc gctacttctg gggtaacctt cccggtatga acaggccgtt ggcatccact
     7741 gtgaatgata agctggagct gcaggagtgt ctggagcatg gcaggatagc caagttcagc
     7801 aaagtgagga ccattactac gaggtcaaac tccataaagc agggcaaaga ccagcatttt
     7861 cctgtcttca tgaatgagaa agaggacatc ttatggtgca ctgaaatgga aagggtattt
     7921 ggtttcccag tccactatac tgacgtgtcc aacatgagcc gcttggcgag gcagagactg
     7981 ctgggccggt catggagcgt gccagtcatc cgccacctct tcgctccgct gaaggagtat
     8041 tttgcgtgtg tggaggtgca caccaaccaa gatccgcttg atgctgaagt tcatacaaac
     8101 caggatcccc tcgatgccga agtccatact aatcaggacc cactggacgc acccaagaaa
     8161 aagcgcaaag tcggcagcgg cgccaccaac ttcagcctgc tgaagcaggc cggcgacgtg
     8221 gaggagaacc ctggacccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     8281 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     8341 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     8401 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     8461 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     8521 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     8581 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     8641 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     8701 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     8761 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     8821 cgcaagcccg gtgcctgacg ggcgcgtctg gaacaatcaa cctctggatt acaaaatttg
     8881 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     8941 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     9001 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     9061 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     9121 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     9181 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     9241 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     9301 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     9361 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     9421 ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct agaaaaacat
     9481 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca
     9541 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact
     9601 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact ggaagggcta
     9661 attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac
     9721 ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact gacctttgga
     9781 tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa taaaggagag
     9841 aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagtg
     9901 ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga gctgcatccg
     9961 gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc tggggacttt
    10021 ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa
    10081 gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct
    10141 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
    10201 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
    10261 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
    10321 acttgcaaag aaatgaatat cagagagtga gaggccttga cattgctagc gtttaccgtc
    10381 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
    10441 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
    10501 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
    10561 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
    10621 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
    10681 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
    10741 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
    10801 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
    10861 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
    10921 ccccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
    10981 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
    11041 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
    11101 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
    11161 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
    11221 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
    11281 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
    11341 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
    11401 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
    11461 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
    11521 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
    11581 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
    11641 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
    11701 aatgataccg cgagatccac gctcaccggc tccagattta tcagcaataa accagccagc
    11761 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
    11821 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
    11881 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
    11941 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
    12001 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
    12061 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
    12121 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
    12181 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
    12241 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
    12301 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
    12361 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
    12421 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
    12481 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
    12541 atttccccga aaagtgccac ctgacgtcga cggatcggga gatcaacttg tttattgcag
    12601 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
    12661 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca
    12721 actggataac tcaagctaac caaaatcatc ccaaacttcc caccccatac cctattacca
    12781 ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa ttattttcat
    12841 tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagt
//
